identify new metabolites of dietary monoterpenes Jarlei Fiamoncini - - PowerPoint PPT Presentation
In silico prediction of metabolism as a tool to identify new metabolites of dietary monoterpenes Jarlei Fiamoncini Food Metabolome and the Metabolism of Food Compounds Food metabolome is the part of the metabolome derived from the digestion and
Limonene Isopentenyl pyrophosphate Dimethylallyl pyrophosphate Geranyl pyrophosphate Geraniol
Limonene Isopentenyl pyrophosphate Dimethylallyl pyrophosphate Geranyl pyrophosphate Geraniol
Antinociceptive Antimicrobial Hypotensive Anti-inflammatory Hypoglycemic (STZ diabetic mice) Antioxidant Antineoplasic Modulators of the activity of ion channels Toxic effects
Both in humans and rats, dietary terpenes reach effective concentrations in plasma within 1 hour Their metabolites are detected in circulation up to 24 hours after intake Topic administration of terpenes is also effective to increase their concentration in plasm
Despite recognized health effects, the metabolism of dietary terpenoids is poorly known Different isomers for each compound make terpenoids analysis very complex.
1,4-Cineole
lime eucalyptus
Citral
lemongrass lemon balm eucalyptus
Fenchone
fennel
Myrcene
hop
Pulegone
mint
Camphene
thyme
Citronellal
lemon balm
Geraniol
lemon grass citronella geranium
Nootkatone
grapefruit
Terpinen-4-ol
juniper
Carvacrol
thyme
Cuminaldehyde
eucalyptus myrrh
Limonene
p-Cymene
cumin thyme
Thymol
thyme
Carvone
caraway spearmint
D-Camphor
camphor tree
Linalool
rosewood coriander
Perillyl alcohol
lavender sage
Tested dietary terpenoids
Caryophyllene
clove cannabis rosemary
1,8-Cineole
eucalyptus
Menthol
mint
Pinene
pine
defining the reactions involved in the the metabolism of dietary monoterpenes
using selected reactions to predict the metabolites of monoterpenes
feeding monoterpenes to rats and collecting metabolites-rich urine
non-targeted high-resolution LC- MS analysis of urine in search of predicted metabolites
http://phytohub.eu/ https://www.lhasalimited.org
Cuminaldehyde metabolite 1 (M7) Cuminaldehyde metabolite 3 (M34) Cuminaldehyde metabolite 2 (M20)
Cuminaldehyde
reduction of aldehydes
hydroxylation of aromatic methine hydroxylation of terminal methyl reduction of aldehydes Cuminaldehyde metabolite 5 (M6)
alcohols Cuminaldehyde metabolite 4 (M140)
aldehydes
M2 M18
M3 M31 M30
Biotransformation Name Phase Enzyme
Compounds that undergo the specific reactions
Allylic Hydroxylation Phase I CYP450
limonene nootkatone geraniol terpinen-4-ol perillyl alcohol linalool
Conjugation of Alkyl Carboxylic Acids with Glycine Phase II ACS, AANAT
geraniol terpinen-4-ol perillyl alcohol
Conjugation of Carboxylic Acids with Glutamine Phase II ACS, AANAT
geraniol
Epoxidation of 1,1,2-Trisubstituted Alkenes Phase I CYP450
limonene geraniol terpinen-4-ol perillyl alcohol linalool
Epoxidation of 1,1-Disubstituted Alkenes Phase I CYP450
limonene nootkatone perillyl alcohol
Epoxidation of Monosubstituted Alkenes Phase I CYP450
linalool
Glucuronidation of Aromatic Alcohols Phase II UGT
thymol
Glucuronidation of Carboxylic Acids Phase II UGT
thymol limonene nootkatone geraniol terpinen-4-ol perillyl alcohol cuminaldehyde linalool menthol
Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols Phase II UGT
thymol limonene nootkatone geraniol terpinen-4-ol perillyl alcohol cuminaldehyde linalool menthol
Hydroxylation of Alkyl Methine Phase I CYP450
nootkatone terpinen-4-ol menthol
Hydroxylation of Aromatic Methine Phase I CYP450
thymol cuminaldehyde
Hydroxylation of Methyl Carbon Adjacent to an Aliphatic Ring Phase I CYP450
nootkatone menthol
Hydroxylation of Methyl Carbon Next to an Aromatic Ring Phase I CYP450
thymol
Hydroxylation of Terminal Methyl Phase I CYP450
thymol terpinen-4-ol cuminaldehyde linalool menthol
Hydroxylation of Unfunctionalised Alicyclic Methylene Phase I CYP450
limonene nootkatone perillyl alcohol menthol
Oxidation of Aldehydes Phase I ALDH
cuminaldehyde
Oxidation of Primary Alcohols Phase I ADH
thymol limonene nootkatone geraniol terpinen-4-ol perillyl alcohol cuminaldehyde linalool menthol
Oxidation of Secondary (Alicyclic) Alcohols Phase I ADH
limonene nootkatone geraniol terpinen-4-ol perillyl alcohol menthol
Reduction of Aldehydes Phase I ALDR
cuminaldehyde
Reduction of Alicyclic Ketones Phase I ADH
menthol
Reduction of alpha,beta-Unsaturated Compounds Phase I abKDBR
nootkatone
Vicinal Diols from Epoxides Phase I EH
limonene nootkatone geraniol perillyl alcohol linalool
1 138 3
Epoxidation of 1,1-Disubstituted Alkenes
LIMONENE
7 4
Vicinal Diols from epoxides
8 9 34 16 24
Oxidation
Secondary (Alicyclic) Alcohols
5
Allylic Hydroxylation Allylic Hydroxylation Allylic Hydroxylation
83
Glucuronidati
and Secondary Aliphatic and Benzylic Alcohols
30 154
Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols
165
Oxidation of Secondary (Alicyclic) Alcohols Vicinal Diols from epoxides Oxidation of Secondary (Alicyclic) Alcohols Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols
196 225
Oxidation of Secondary (Alicyclic) Alcohols
40
Glucuronidation
Secondary Aliphatic and Benzylic Alcohols
45
Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols
255 245
Oxidation
Alcohols
62
Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols
64 69 68
Oxidation of Secondary (Alicyclic) Alcohols Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols Epoxidation of 1,1,2- Trisubstituted Alkenes Oxidation
Alcohols Allylic Hydroxylation Glucuronidation of Primary and Secondary Aliphatic and Benzylic Alcohols
Oxidation
Alcohols
Hydroxylation of Unfunctionalised Alicyclic Methylene
Bio BioTransformer
Uni niversity ty of
rta
8 days on AIN-93 diet wash-out period 2 male, wistar rats 2 female, wistar rats
Urine sampling start
5 days on AIN-93 supplemented with 0,05% terpenes (15mg /day) 2 male, wistar rats 2 female, wistar rats
Urine sampling
5 cycles – same rats were exposed to different food compounds
PhytoHub 2 PhytoHub 1 PhytoHub 3 PhytoHub 4
356,111 340,116 194,058 194,058 178,063 166,099 164,084 148,089
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Known metabolites Predicted metabolites Experimental data on rat metabolites
Literature & Databases
The structures in the chromatogram were not yet confirmed. They have the same mass as the assigned peaks and are therefore used as examples. There are other structures predicted with same molecular mass.
The structures in the chromatogram were not yet confirmed. They have the same mass as the assigned peaks and are therefore used as examples. There are other structures predicted with same molecular mass.
Considering the selected 22 biotransformations, more than 1500 metabolites were predicted from the 23 tested terpenoids. The predicted metabolites were helpful for the annotation of the peaks detected after the rats were exposed to the terpenoids. Next step is to validate the hypothetical structures of known and new metabolites using qToF MS/MS. The knowledge generated is being used to improve in silico prediction tools (BioTransformer) The generated data will be made available in food compounds databases (PhytoHub, HMDB)
laudine Manach